Target Protein: Dihydrofolate Reductase (Bacillus anthracis)
E.C. Number: 1.5.1.3
Gene #: 3361714
Gene symbol: pxo1_110
Genomic Sequence: NC_001496.1

PDB ID: 2QK8
Protein ID: Q81R22
Organism: Bacillus anthracis

Etiological Risk Group: Category B Priority Pathogens -Bacillus anthracis

Disease:
Anthrax is a zoonotic disease caused by Bacillus anthracis, an aerobic, gram-positive spore-forming bacterium. Bacillus anthracis primarily affects herbivores; though most warmed-blooded mammals may be susceptible [1], including humans. Anthrax is an ancient disease that has caused losses of livestock and wildlife populations prior to and throughout the 20th century and remains enzootic with seasonal variations in many parts of the world [2, 3].
http://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0004689#abstract0

Homology: Bacillus cereus, Bacillus thuringiensis, and Bacillus mycoides
Essentiality: Bacillus anthracis-derived nitric oxide is essential for pathogen virulence and survival of macrophages.
Phagocytes generate nitric oxide (NO) and other reactive oxygen and nitrogen species in large quantities to combat infecting bacteria. Here, we report the surprising observation that in vivo survival of a notorious pathogen—Bacillus anthracis—critically depends on its own NO-synthase (bNOS) activity. Anthrax spores (Sterne strain) deficient in bNOS lose their virulence in an A/J mouse model of systemic infection and exhibit severely compromised survival when germinating within macrophages.

http://www.pnas.org/content/105/3/1009

Druggable Target: One possible candidate is dihydrofolate reductase (DHFR), a biosynthetic enzyme necessary for anthrax pathogenicity. We determined the crystal structure of DHFR from B. anthracis (baDHFR) in complex with methotrexate.

Protein Image:

2qk8.jpg
Figure 1. PyMol image crystal structure of the anthranx drug target, Bacillus anthracis dihydrofolate reductase

Assayable Enzyme Target:
1.5.1.3 Dihydrofolate Reductase
https://www.sigmaaldrich.com/life-science/metabolomics/enzyme-explorer/learning-center/assay-library/ec-number.html

Structure (PDB or Homology model)
PDB # or closest PDB entry if using homology model: 3HMC
For Homology Model option: N/A
Expression System: Escherichia coli BL21(DE3)
Inhibitors: n/a


Pairwise alignment of your BLASTP search in NCBI against the PDB--100% aligned
Capture (2).PNG
https://blast.ncbi.nlm.nih.gov/Blast.cgi#alnHdr_446669140
Gene Availability:
http://www.uniprot.org/proteomes/UP000000427
https://www.ncbi.nlm.nih.gov/gene/3361714
Protein sequence:
>2QK8:A|PDBID|CHAIN|SEQUENCE

MIVSFMVAMDENRVIGKDNNLPWRLPSELQYVKKTTMGHPLIMGRKNYEAIGRPLPGRRNIIVTRNEGYHVEGCEVAHSV

EEVFELCKNEEEIFIFGGAQIYDLFLPYVDKLYITKIHHAFEGDTFFPEMDMTNWKEVFVEKGLTDEKNPYTYYYHVYEK

QQ
length of your protein in Amino Acids: 162 aa
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 19.124 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 1.439

TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
TMPRED.1664.2779.gif

CDS Gene Sequence:

<span style="background-color: #ffffff; font-family: monospace,serif;">>NC_001496.1:c135455-133161 Bacillus anthracis virulence plasmid PX01, complete sequence
ATGAAAAAACGAAAAGTGTTAATACCATTAATGGCATTGTCTACGATATTAGTTTCAAGCACAGGTAATT
TAGAGGTGATTCAGGCAGAAGTTAAACAGGAGAACCGGTTATTAAATGAATCAGAATCAAGTTCCCAGGG
GTTACTAGGATACTATTTTAGTGATTTGAATTTTCAAGCACCCATGGTGGTTACCTCTTCTACTACAGGG
GATTTATCTATTCCTAGTTCTGAGTTAGAAAATATTCCATCGGAAAACCAATATTTTCAATCTGCTATTT
GGTCAGGATTTATCAAAGTTAAGAAGAGTGATGAATATACATTTGCTACTTCCGCTGATAATCATGTAAC
AATGTGGGTAGATGACCAAGAAGTGATTAATAAAGCTTCTAATTCTAACAAAATCAGATTAGAAAAAGGA
AGATTATATCAAATAAAAATTCAATATCAACGAGAAAATCCTACTGAAAAAGGATTGGATTTCAAGTTGT
ACTGGACCGATTCTCAAAATAAAAAAGAAGTGATTTCTAGTGATAACTTACAATTGCCAGAATTAAAACA
AAAATCTTCGAACTCAAGAAAAAAGCGAAGTACAAGTGCTGGACCTACGGTTCCAGACCGTGACAATGAT
GGAATCCCTGATTCATTAGAGGTAGAAGGATATACGGTTGATGTCAAAAATAAAAGAACTTTTCTTTCAC
CATGGATTTCTAATATTCATGAAAAGAAAGGATTAACCAAATATAAATCATCTCCTGAAAAATGGAGCAC
GGCTTCTGATCCGTACAGTGATTTCGAAAAGGTTACAGGACGGATTGATAAGAATGTATCACCAGAGGCA
AGACACCCCCTTGTGGCAGCTTATCCGATTGTACATGTAGATATGGAGAATATTATTCTCTCAAAAAATG
AGGATCAATCCACACAGAATACTGATAGTCAAACGAGAACAATAAGTAAAAATACTTCTACAAGTAGGAC
ACATACTAGTGAAGTACATGGAAATGCAGAAGTGCATGCGTCGTTCTTTGATATTGGTGGGAGTGTATCT
GCAGGATTTAGTAATTCGAATTCAAGTACGGTCGCAATTGATCATTCACTATCTCTAGCAGGGGAAAGAA
CTTGGGCTGAAACAATGGGTTTAAATACCGCTGATACAGCAAGATTAAATGCCAATATTAGATATGTAAA
TACTGGGACGGCTCCAATCTACAACGTGTTACCAACGACTTCGTTAGTGTTAGGAAAAAATCAAACACTC
GCGACAATTAAAGCTAAGGAAAACCAATTAAGTCAAATACTTGCACCTAATAATTATTATCCTTCTAAAA
ACTTGGCGCCAATCGCATTAAATGCACAAGACGATTTCAGTTCTACTCCAATTACAATGAATTACAATCA
ATTTCTTGAGTTAGAAAAAACGAAACAATTAAGATTAGATACGGATCAAGTATATGGGAATATAGCAACA
TACAATTTTGAAAATGGAAGAGTGAGGGTGGATACAGGCTCGAACTGGAGTGAAGTGTTACCGCAAATTC
AAGAAACAACTGCACGTATCATTTTTAATGGAAAAGATTTAAATCTGGTAGAAAGGCGGATAGCGGCGGT
TAATCCTAGTGATCCATTAGAAACGACTAAACCGGATATGACATTAAAAGAAGCCCTTAAAATAGCATTT
GGATTTAACGAACCGAATGGAAACTTACAATATCAAGGGAAAGACATAACCGAATTTGATTTTAATTTCG
ATCAACAAACATCTCAAAATATCAAGAATCAGTTAGCGGAATTAAACGCAACTAACATATATACTGTATT
AGATAAAATCAAATTAAATGCAAAAATGAATATTTTAATAAGAGATAAACGTTTTCATTATGATAGAAAT
AACATAGCAGTTGGGGCGGATGAGTCAGTAGTTAAGGAGGCTCATAGAGAAGTAATTAATTCGTCAACAG
AGGGATTATTGTTAAATATTGATAAGGATATAAGAAAAATATTATCAGGTTATATTGTAGAAATTGAAGA
TACTGAAGGGCTTAAAGAAGTTATAAATGACAGATATGATATGTTGAATATTTCTAGTTTACGGCAAGAT
GGAAAAACATTTATAGATTTTAAAAAATATAATGATAAATTACCGTTATATATAAGTAATCCCAATTATA
AGGTAAATGTATATGCTGTTACTAAAGAAAACACTATTATTAATCCTAGTGAGAATGGGGATACTAGTAC
CAACGGGATCAAGAAAATTTTAATCTTTTCTAAAAAAGGCTATGAGATAGGATAA
 
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